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Proteomic, transcriptomic and functional characterization of human astrocyte‐derived extracellular vesicles upon inflammatory activation
Author(s) -
You Yang,
Borgmann Kathleen,
Edara VenkataViswa,
Stacy Satomi,
Ghorpade Anuja,
Ikezu Tsuneya
Publication year - 2020
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1002/alz.039585
Subject(s) - microbiology and biotechnology , biology , microvesicles , proteomics , astrocyte , cell signaling , exosome , transcriptome , proteome , quantitative proteomics , signal transduction , microrna , biochemistry , gene expression , central nervous system , neuroscience , gene
Background Astrocytes in the central nervous system provide supportive neural functions and mediate inflammatory responses from microglia. Increasing evidence supports their critical roles in regulating brain homeostasis in response to pro‐inflammatory factors such as cytokines and pathogen/damage‐associated molecular pattern molecules in infectious and neurodegenerative diseases. However, the underlying mechanisms of the trans‐cellular communication is still unclear. Extracellular vesicles (EVs) can transfer a large diversity of molecules such as lipids, nucleic acids and proteins for cellular communications. The purpose of this study is to characterize the EVs cargo proteins derived from human primary astrocytes (AstrEVs) under both physiological and pathophysiological conditions. Methods We performed a quantitative proteomic and transcriptomic analysis of exosomes purified from human primary astrocytes with or without interleukin 1‐β (IL1‐β) stimulation in vitro. Exosome‐enriched fractions were purified by size‐exclusion columns. The total proteins isolated from the EVs were run on 1D SDS‐PAGE and mass spectrometry. miRNA was isolated from EVs and subjected to Affymetrix miRNA 4. 0 Array. The data are subjected to bioinformatic analysis and validation for select molecules. Human astrocyte‐derived EVs were tested for their effect on neuronal differentiation and maturation by live confocal imaging and multi‐electrode array. Results Label‐free quantitative proteomic profiling revealed a notable up‐regulation of proteins including actin‐associated molecules, integrins and major histocompatibility complex in IL‐1β‐AstrEVs compared to CTL‐AstrEVs, which were involved in cellular metabolism and organization, cellular communication and inflammatory response. They were also significantly associated with the increased signaling of pathogenic molecules such as APP and tau. IL‐1β‐AstrEVs show increased uptake by primary murine cortical neurons compared to CTL‐AstrEVs, which was partially suppressed by RGD peptide, a pan‐integrin inhibitor. Additionally, treatment of neurons with IL‐1β‐AstrEVs also reduced neurite outgrowth, branching and neuronal firing. Conclusions These findings provide insight for the molecular mechanism of the AstrEVs’ effects on neural uptake, neural differentiation and maturation, and its alteration in inflammatory conditions, which may be relevant to neuroinflammation‐mediated Alzheimer’s disease progression.